Au400 analyzer

A serum alanine aminotransferase (ALT) Beckman Coulter test kit (OSR 6107) and aspartate aminotransferase (AST) were performed on the sera harvested at sacrifice from experimental mice according to the manufacturer’s protocol and analyzed using a Beckman Coulter AU400 analyzer by the Clinical Biochemistry Unit, Women’s Hospital, Edgbaston, Birmingham, U.K.

At 16 weeks, blood was collected by puncture of the retro-orbital sinus under terminal anaesthesia within 1−3 hours of lights on and collected into ethylenediaminetetraacetic acid-coated tubes (Kabe Labortechnik, Numbrecht, Germany) for hematology (Scil Vetabc, Montpellier, France) and into heparinized tubes (Kabe Labortechnik) for plasma preparation. A total of 26 parameters were determined from plasma using an Olympus AU400 analyzer (Beckman Coulter, High Wycombe, UK). Insulin and erythropoietin were determined using a Meso Scale Discovery array (Rockville, MD) and interleukin 6 was measured by enzyme-linked immunosorbent assay (eBioscience, Hatfield, UK).

Fasting blood samples were collected by qualified nurses in the Clinical Laboratory unit at the Department of Sport and Health. Spanish Agency for Health Protection in Sport (AEPSAD). Complete blood counts were determined from Vacutainer^TM EDTA tubes (Becton Dickinson, Franklin Lakes, NY, USA) by standardized clinical laboratory procedures using an automated hematology analyzer (ADVIA 120; Siemens Healthcare Diagnostics Inc., Deerfield, IL, USA) within 4h after extraction. For serum separation, blood samples were collected in Vacutainer^TM gel tubes (No additive (Z) [40 ,41 (link),42 (link)] (Becton Dickinson, Franklin Lakes, NY, USA). Samples were stored at room temperature for at least 15 min to allow clot formation and then immediately centrifuged for 15 min at 4000 rpm and 10 °C. Serum samples were aliquoted and analyzed to assess basic biochemistry in an AU400 analyzer (Beckman Coulter Inc., Brea, CA, USA) and stored at −80 °C for later analysis.